On-chip valves can simplify a microfluidic chip and make it easy to operate. However, most on-chip valves already reported still need complicated manufacture and sophisticated supporting devices. In this work, we present a straightforward on-chip valve, which can be serially connected, to form an on-chip pump. The liquid can horizontally flow one way by the regular deformations of flexure strips in the two valves at both sides of the chamber under pressure changes in microchannels generated by repeated vertical movements of linear actuators. The volume of this system including the chip and the supporting device is 0.65 cubic decimeters, which is much smaller than that of reported systems with a volume of at least 12 cubic decimeters, and the weight of this system is only 0.56 kg, making it possible for point-of-care testing. We carry out an immunoassay of folic acid on chip, and the results show satisfactory reproducibility with acceptable coefficients of variation. We determine 163 clinical human serum samples for folic acid. Furthermore, we detect transferrin, cobalamin and folic acid simultaneously on one chip with both sandwich and competitive binding immunoassay methods. We anticipate that this on-chip valve and pump can be applied in immunoassays and other biosensing applications.